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Related Experiment Videos

Apical membrane permeability of MDCK cells

R L Rivers1, J A McAteer, J L Clendenon

  • 1Department of Cell Biology and Anatomy, Medical University of South Carolina, Charleston 29425-2204, USA.

The American Journal of Physiology
|July 11, 1996
PubMed
Summary

This study precisely measured water and solute permeability of Madin-Darby canine kidney (MDCK) cell apical membranes using a novel cyst model. Results reveal high osmotic water permeability (Pf) and solute transport characteristics, crucial for understanding epithelial function.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Renal Physiology

Background:

  • Madin-Darby canine kidney (MDCK) C12 cells are a widely used model for epithelial research.
  • Understanding apical membrane permeability is crucial for epithelial transport studies.
  • Previous methods for measuring membrane permeability often used isolated vesicles, not intact cells.

Purpose of the Study:

  • To determine the osmotic water permeability (Pf) and nonelectrolyte permeability of the apical membrane of MDCK C12 cell cysts.
  • To develop and validate a novel method for measuring membrane permeability in intact epithelial cells.
  • To investigate the influence of cell number and specific substances on membrane permeability.

Main Methods:

  • Cultured MDCK C12 cells as apical-out cysts, with the apical membrane exposed to the medium.

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  • Applied instantaneous osmotic challenges to cysts and recorded volume changes via automated image analysis.
  • Calculated Pf and solute permeabilities from volume change rates and fitted data to determine kinetic parameters.

    • Main Results:

    • A precise average Pf of 2.4 +/- 0.1 micron/s was determined for the MDCK apical membrane.
    • Amphotericin B increased Pf, while forskolin had no significant effect.
    • Urea permeability was negligible, but acetamide and formamide permeabilities correlated with their oil-water partition coefficients.
    • Paracellular water flow was found to be insignificant in multi-cell cysts.

    Conclusions:

    • The apical-out MDCK cyst model combined with automated image analysis provides a highly precise method for determining epithelial apical membrane permeability.
    • The measured Pf and solute permeabilities are consistent with those of water-tight epithelial apical membranes.
    • This technique offers a valuable tool for investigating epithelial transport mechanisms in a more physiologically relevant context.